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E6基因产物对非洲爪蟾卵中1型牛乳头瘤病毒DNA复制的选择性增强作用。

Selective enhancement of bovine papillomavirus type 1 DNA replication in Xenopus laevis eggs by the E6 gene product.

作者信息

Romanczuk H, Wormington W M

机构信息

Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02254.

出版信息

Mol Cell Biol. 1989 Feb;9(2):406-14. doi: 10.1128/mcb.9.2.406-414.1989.

DOI:10.1128/mcb.9.2.406-414.1989
PMID:2540419
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC362615/
Abstract

Genetic analyses of bovine papillomavirus type 1 (BPV-1) DNA in transformed mammalian cells have indicated that the E6 gene product is essential for the establishment and maintenance of a high plasmid copy number. In order to analyze the direct effect of the E6 protein on the replication of a BPV-1-derived plasmid, a cDNA containing the BPV-1 E6 open reading frame was subcloned into an SP6 vector for the in vitro synthesis of the corresponding mRNA. The SP6 E6 mRNA was injected into Xenopus laevis oocytes to determine the subcellular localization of the E6 gene product and to analyze the effect of the protein on BPV-1 DNA replication. SP6 E6 mRNA microinjected into stage VI oocytes was translated into a 15.5-kilodalton protein that was specifically immunoprecipitated by antibodies directed against the E6 gene product. The E6 protein preferentially accumulated in oocyte nuclei, a localization which is consistent with the replicative functions in which it has been implicated. The expression of E6 in replication-competent mature oocytes selectively enhanced the replication of a BPV-derived plasmid, indicating a direct role for this gene product in the control of BPV-1 DNA replication.

摘要

对转化的哺乳动物细胞中1型牛乳头瘤病毒（BPV-1）DNA的遗传分析表明，E6基因产物对于高质粒拷贝数的建立和维持至关重要。为了分析E6蛋白对BPV-1衍生质粒复制的直接影响，将包含BPV-1 E6开放阅读框的cDNA亚克隆到SP6载体中，用于体外合成相应的mRNA。将SP6 E6 mRNA注射到非洲爪蟾卵母细胞中，以确定E6基因产物的亚细胞定位，并分析该蛋白对BPV-1 DNA复制的影响。注射到VI期卵母细胞中的SP6 E6 mRNA被翻译成一种15.5千道尔顿的蛋白质，该蛋白质被针对E6基因产物的抗体特异性免疫沉淀。E6蛋白优先积累在卵母细胞核中，这种定位与其所涉及的复制功能一致。在具有复制能力的成熟卵母细胞中E6的表达选择性地增强了BPV衍生质粒的复制，表明该基因产物在BPV-1 DNA复制的控制中起直接作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/2e64bb70605d/molcellb00050-0067-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/ae8702f120aa/molcellb00050-0062-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/34d4a073c069/molcellb00050-0063-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/7b582b35b167/molcellb00050-0063-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/340029901dc0/molcellb00050-0064-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/9e2050cce272/molcellb00050-0064-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/4fc10c95e44e/molcellb00050-0065-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/64036138fbcf/molcellb00050-0066-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/2e64bb70605d/molcellb00050-0067-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/ae8702f120aa/molcellb00050-0062-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/34d4a073c069/molcellb00050-0063-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/7b582b35b167/molcellb00050-0063-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/340029901dc0/molcellb00050-0064-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/9e2050cce272/molcellb00050-0064-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/4fc10c95e44e/molcellb00050-0065-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/64036138fbcf/molcellb00050-0066-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9001/362615/2e64bb70605d/molcellb00050-0067-a.jpg

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